This proposal describes the creation of a Neuroproteomics Center that will provide proteomics and bioinformatics technologies to the UIUC neuroscience community while simultaneously advancing the performance of state-of-the-art proteomics technologies to new levels of performance, especially for the study of addiction mechanisms in the central nervous system. This Center integrates research groups with expertise in the fields of analytical chemistry and bioinformatics with those in biological and behavioral neuroscience in a unified, directed approach to discover the intricacies of intercellular signaling. Specifically, the Center will be built around the overarching theme of cell-cell signaling, which will be complemented by two technology development thrusts. Extracellular signaling peptides and proteins - neuropeptides, trophic factors, cytokines, and hormones - represent a critical part of the cell proteome that has been implicated in almost all aspects of organism function, including learning and memory. But measuring the complex array of such proteins and protein-derived factors, especially when found only in small regions of the brain, has been difficult because of a lack of approaches that allow small samples to be probed with the required chemical information content. Because these signaling molecules tend to be among the compounds that undergo the most post-translational modification, are active at the lowest levels, and are spatially localized to distinct physical regions, the development of ultra small sampling and sophisticated detection technologies is required. Accomplishment of these Aims will enable enhanced comparison in various animal models of genomic and functional data with proteomic data, and will expand the concept of directed proteomics by pushing the limits on small volume sampling, purification, detection, and characterization. The Center is divided into four core groups: the Sampling and Separation Core, the Protein Identification Core, and the Bioinformatics Core, plus the Administrative Core, with each described in separate sections of the proposal. The major biological users include five laboratories at UIUC: David Clayton, Martha Gillette, Paul Gold, William Greenough, and Gene Robinson, and two outside laboratories: Howard Gutstein of the Anderson Cancer Center and Leonid Moroz at the University of Florida.